China
Africa
Explore chapters and articles related to this topic
Use of Fluorescence as a Voltage Indicator in Mononuclear Cells*
Published in Richard C. Niemtzow, Transmembrane Potentials and Characteristics of Immune and Tumor Cell, 2020
Jeffrey L. Rossio, Richard C. Niemtzow
A second class of dyes, represented by acridine orange and mithramycin, bind to nucleic acids. Since one correlate of cell activation is change in the nuclear morphology (representing, perhaps, more transcriptive activity), these dyes do present a rough correlation with activation. But these changes occur slowly and often are not predictably consistent among various cell types.
Liver Microcirculation
Published in John H. Barker, Gary L. Anderson, Michael D. Menger, Clinically Applied Microcirculation Research, 2019
Acridine orange (1 µmol/kg, Sigma, St. Louis) is given as fluorescence marker of leukocytes, allowing assessment of sinusoidal perfusion and leukocyte endothelial interactions.69 For determination of red blood cell velocity, ex vivo FITC-labeled erythrocytes are used, as originally described by Zimmerhackl et al. in a modified technique.70 The labeled cells are stored at +4°C after addition of citrate-phosphate-dextrose (1.4:10) up to 5 days. Two min prior to liver microscopy, 0.05 ml red cells, 1:1 diluted with normal saline, are injected into the isogenic recipient animals. For investigation of macrophage function, fluorescence-labeled latex particles of approximately 1-µm diameter (Polysciences) are injected intravenously, as described earlier.39,67
New methodology for microbiological quality assurance
Published in R. M. Baird, S. F. Bloomfield, Microbial quality assurance in cosmetics, toiletries and non-sterile Pharmaceuticals, 2017
This technique depends on the observation that viable acridine orange-stained bacteria tend to fluoresce orange/red. Micro-organisms concentrated by filtration from the original sample are stained with acridine orange and viable cells counted under a fluorescent microscope. The method can detect greater than 103–104 micro-organisms ml−1 of product (Pettipher and Rodrigues 1981) although this sensitivity greatly depends on the quantity of product that can be filtered. The direct epifluorescent filtration technique (DEFT) has a number of distinct advantages over many other methods (Fung 1991). Firstly, it is a truly rapid technique, the method taking less than 1 h for sample analysis. Secondly, it is a quantitative technique (Rodrigues and Kroll 1985) although there may be problems in differentiating viable from non-viable cells in some samples (Rodrigues and Kroll 1986). The DEFT has been investigated for milk analysis (Pettipher et al. 1980), milk product analysis (Pettipher and Rodrigues 1981) and food analysis (Pettipher and Rodrigues 1982). The DEFT has also been evaluated for the detection of bacterial contaminants in intravenous fluids where detection down to 25 organisms ml−1 was obtained by increasing the sample volume filtered (Denyer and Ward 1983). Recently a method has been developed to overcome the problem of non-viable cells staining orange/red. This involved filtration of the sample followed by a short incubation to allow microcolonies to form. The sample was stained and the viable microcolonies were then counted (Newby 1991).
Impact of bee venom and melittin on apoptosis and biotransformation in colorectal carcinoma cell lines
Published in Toxin Reviews, 2021
Danijela D. Nikodijević, Milena G. Milutinović, Danijela M. Cvetković, Maja Đ. Ćupurdija, Milena M. Jovanović, Ivan V. Mrkić, Marija Đ. Jankulović-Gavrović, Snežana D. Marković
Phosphate-buffered saline (PBS) and Dulbecco’s modified Eagle medium (DMEM) were obtained from GIBCO (Invitrogen, Carlsbad, CA). Ethidium bromide (EB), 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT), absolute ethanol and chloroform (molecular biology grade) were obtained from SERVA (Heidelberg, Germany). Acridine orange (AO) was obtained from Acros Organics (Morris Plains, NJ). qPCR Kit (Luna Universal qPCR Master Mix Kit) was from BioLabs, New England (Ipswich, MA). Nuclease-free water and TRIzol were from Ambion (Austin, TX). The secondary antibody conjugated with Cy3, diamidino-2-phenylindole (DAPI) and kit for translating RNA into complementary DNA (high-capacity cDNA reverse transcription kits) from Thermo Scientific (Waltham, MA). Polyvinyl alcohol mounting medium was obtained from Fluka Analytical (Buchs, Switzerland). All solvents and chemicals were of analytical grade. Melittin (dry powder) was obtained from Sigma-Aldrich (St. Louis, MO).
Investigation of in vivo unscheduled DNA synthesis in rabbit corneas following instillation of genotoxic agents
Published in Cutaneous and Ocular Toxicology, 2021
Haruna Tahara, Shingo Nemoto, Yoshinori Yamagiwa, Yu Haranosono, Masaaki Kurata
Table 2 summarises the histopathological findings in the corneal epithelium following treatment with test compounds and controls in Experiment 2. No histopathological changes in the corneal epithelium were observed in the negative control (physiological saline)- or vehicle control (DMSO)-treated eyes (Figure 1(d)). In the paraquat-treated eyes, no histopathological changes were observed in the corneal epithelium treated with a concentration of 5% paraquat. Very slight hypertrophy of the corneal epithelial cells was observed at paraquat concentrations of 10% or above. In addition, very slight degeneration/necrosis of superficial cells was observed in some eyes treated with concentrations of 20% and 40% paraquat (Figure 1(e)). There was no tissue sample unsuitable for counting the number of SLCs in any concentration. In the acridine orange-treated eyes, very slight hypertrophy of the corneal epithelial cells, very slight to slight pale-colored cytoplasm and vacuolar degeneration of epithelial cells were observed at acridine orange concentrations of 2.5% or above. In addition, very slight degeneration/necrosis of superficial cells was observed at acridine orange concentrations of 2.5% or above (Figure 1(f)). The number of SLCs could be counted in all tissue samples. In the positive control (1 J/cm2 UV irradiation)-treated eyes, moderate degeneration/necrosis of the corneal epithelial cells was observed in the surface layer.
Anticarcinogenic potential of gold nanoparticles synthesized from Trichosanthes kirilowii in colon cancer cells through the induction of apoptotic pathway
Published in Artificial Cells, Nanomedicine, and Biotechnology, 2019
Xiaodong Han, Xiaojia Jiang, Lanjie Guo, Yongxin Wang, Vishnu Priya Veeraraghavan, Surapaneni Krishna Mohan, Zhigang Wang, Dandan Cao
Moreover, we carried out AO/EB staining method to understand the therapeutic effect of AuNPs on morphological alerations in HCT-116 cells after the treatment. Using a fluorescence microscope in acridine orange ethidium bromide assay, AO/EB staining method illustrated that untreated cells appeared homogeneously green whereas early hours apoptotic cell was yellowish green or yellow colour. Acridine orange was used in conjunction with ethidium bromide to distinguish between viable, necrotic and apoptotic cells [36]. Figure 7 shows control HCT-116 cells as well as necrotic and apoptotic cells after the staining of Acridine orange/ethidium bromide. The HCT-116 control cells did not receive AuNPs treatment represented in green color (live cells) and AuNPs treated cells showed late apoptotic stage in orange colour (dead cells) with chromatin clumping and condensation. The cell-cycle analysis is being beneficial, paricularlly for the sensitization of malignant cells to specific cancer treatment method [37,38]. AuNPs efficiently arrest the cell-cycle progress specifically at the phase of G0/G1and it consequently leads to apoptosis [39]. In the current study also, AuNPs (15 and 20 μg/ml) treated cells showed apoptoic cells which was achieved by the arrest in the phase of G1 (Figure 8). Our results also suggested that AuNPs competently arrest the cell-cycle progress by the induction of apoptosis in HCT-116 cells.